调节性B细胞在接受脱敏治疗过敏性哮喘患儿中的变化及意义

Alternation and its significance of regulatory B cells in allergic asthmatic children with mite-specific immunotherapy

目的探讨过敏性哮喘患儿接受螨虫过敏原特异性免疫治疗(allergen-specific immunotherapy,AIT)前后外周血调节性B细胞(regulatory B cells,Breg)的改变及可能意义。方法接受AIT过敏性哮喘患儿20例,在治疗开始前及剂量递增阶段结束后分别取外周血。采用流式细胞术检测CD19+CD24highCD27+Breg、Th1、Th2、Th17、Treg细胞比例及Breg相关表面分子CD80、CD86、CD1d表达;real-time实时荧光定量PCR检测IL-10、TGF-β、IFNG、T-bet、IL-4、GATA3、IL-17A、RORγt、Foxp3 mRNA水平;ELISA检测IL-10、TGF-β蛋白浓度。结果在AIT剂量递增阶段结束后,CD19+CD24highCD27+Breg比例[(7.53±4.37)vs.(11.46±3.95),P<0.05],IL-10 mRNA水平[(0.87±1.07)×10-1 vs.(1.50±0.78)×10-1,P<0.05],IL-10蛋白水平[(4.87±2.42)vs.(7.07±2.55)]pg/ml,P<0.05],TGF-β蛋白水平[(469.18±182.14)vs.(560.16±163.98)]pg/mL,P<0.05]均较治疗前显著升高;T-bet[(6.30±5.55)×10-2 vs.(3.43±1.63)×10-2]、GATA3[(16.56±11.69)×10-2 vs.(8.18±6.15)×10-2]、RORrt[(19.89±24.22)×10-2 vs.(2.90±5.44)×10-2]mRNA较治疗前均下降(P<0.05)。结论过敏性哮喘患儿接受螨虫AIT治疗,在剂量递增阶段以Breg数量升高及IL-10介导的免疫调节为主,Treg、TGF-β在此阶段可能并未发挥主要作用。

Objective To investigate the alternation and its potential significance of regulatory B cells(Bregs)in allergic asthmatic children with mite specific immunotherapy(AIT).Methods Twenty allergic asthmatic children with AIT were recruited.Peripheral blood samples were collected before initiation and at the end of build-up stage of AIT.(1)The proportions of CD19+CD24highCD27+Breg,Th1,Th2,Th17,Treg cells and expression levels of Breg cell surface molecule CD80,CD86,and CD1d were measured by using flow cytometry;(2)IL-10,TGF-β,IFNG,T-bet,IL-4,GATA3,IL-17A,RORγt,FOXP3 mRNA levels were detected by using real-time fluorescent quantitative PCR;(3)The concentrations of IL-10 and TGF-βwere detected basing on ELISA method.Results The proportion of CD19+CD24highCD27+Breg increased[(7.53±4.37)vs.(11.46±3.95),P<0.05],the expression level of IL-10 mRNA in CD19+B cells increased[(0.87±1.07)×10-1 vs.(1.50±0.78)×10-1,P<0.05],the level of IL-10[(4.87±2.42)vs.(7.07±2.55)]pg/mL,P<0.05]and TGF-β[(469.18±182.14)vs.(560.16±163.98)]pg/mL,P<0.05]in peripheral plasma were significantly increased at end of build-up stage,compared to baseline.The mRNA levels of T-bet[(6.30±5.55)×10-2 vs.(3.43±1.63)×10-2]、GATA3[(16.56±11.69)×10-2 vs.(8.18±6.15)×10-2]and RORrt[(19.89±24.22)×10-2 vs.(2.90±5.44)×10-2]decreased,compared to pre-AIT(P<0.05).ConclusionsFor allergic asthmatic children with mite AIT,the number of Bregs and IL-10-mediated immune regulation were major up-regulated factors in build-up stage,while Treg and TGF-βmight not play a key role in this stage.